Experimental Infection of Rabbits with Newly Excysted Metacercariae of Japanese Fasciola Sp. and American Fasciola Hepatica by Portal Vein Route

日本産およびアメリカ産脱嚢幼肝蛭を家兎の門脈内に移入・感染させ,各種変化について検索した。幼肝蛭は感染後3時間で肝実質内に認められ,その後感染経過に伴い肝病変は漸次増大した。アメリカ産肝蛭では胆管への定着と虫卵排出も認められた。また,日本産肝蛭についても感染後虫体は漸次成長することが認められた。沈降抗体の出現時期は,感染後21-28日であった。幼肝蛭は血行によって肝臓に達してもよく発育し,感染が成立することが明らかとなった。 / Rabbits were experimentally infected with newly excysted metacercariae of the Japanese Fasciola sp. and the American Fasciola hepatica by portal vein route. At early stages of the infection, histopathological changes of the liver of rabbits infected with either F. sp. or F. hepatica were tract lesions characterized by haemorrhages, necrosis, cellular infiltration, and so on. The tract lesions grew larger and more numerous with the progress of the infection. At the 77th day of infection, increases of connective tissue around the intrahepatic bile ducts and in thickness of the wall of the bile ducts were noted. Precipitating antibodies were first detected at the 28th and 21st days of infection in sera of the rabbits infected with F. sp. and F. hepatica, respectively. Fluke eggs were first detected at the 63rd day in feces of the rabbits infected with F. hepatica, and were not detected even at the 77th day in the case of F. sp. Fluke eggs recovered from the infected rabbits were measured the maximum length of 15.9mm and 16.2mm, for F. sp. and F. hepatica, respectively. The infection of rabbits with F. sp. and F. hepatica by the vein route was successful

Improved method of expression cloning in a host cell

The invention relates to a promoter DNA sequence highly suited in an improved expression cloning method for isolation of DNA sequences comprising a DNA sequence encoding a protein of interest in a host cell and to the improved expression cloning method wherein use is made of this promoter. The isolated DNA sequences are useful in processes for producing a protein of interest

Peroxicretion: a novel secretion pathway in the eukaryotic cell

Background: Enzyme production in microbial cells has been limited to secreted enzymes or intracellular enzymes followed by expensive down stream processing. Extracellular enzymes consists mainly of hydrolases while intracellular enzymes exhibit a much broader diversity. If these intracellular enzymes could be secreted by the cell the potential of industrial applications of enzymes would be enlarged. Therefore a novel secretion pathway for intracellular proteins was developed, using peroxisomes as secretion vesicles. Results: Peroxisomes were decorated with a Golgi derived v-SNARE using a peroxisomal membrane protein as an anchor. This allowed the peroxisomes to fuse with the plasma membrane. Intracellular proteins were transported into the peroxisomes by adding a peroxisomal import signal (SKL tag). The proteins which were imported in the peroxisomes, were released into the extracellular space through this artificial secretion pathway which was designated peroxicretion. This concept was supported by electron microscopy studies. Conclusion: Our results demonstrate that it is possible to reroute the intracellular trafficking of vesicles by changing the localisation of SNARE molecules, this approach can be used in in vivo biological studies to clarify the different control mechanisms regulating intracellular membrane trafficking. In addition we demonstrate peroxicretion of a diverse set of intracellular proteins. Therefore, we anticipate that the concept of peroxicretion may revolutionize the production of intracellular proteins from fungi and other microbial cells, as well as from mammalian cells.

Global value perceptions : the legitimising functions of western representations of democracy

This paper argues that a fundamental antagonism between democracy and nondemocracy organises lay thinking on global issues. We review key findings of a long-standing experimental research programme that examined the "Democracy-as-value" hypothesis across a variety of political and social contexts. This hypothesis contends that democracy is an ideological belief system that provides value to democratic individuals, groups, and institutions and thereby grants legitimacy to their actions. Based on procedural justice theories and social representations theory, we contend that western lay perceivers associate democracy with procedural equality and individual autonomy, whereas nondemocracy is associated with ingroup hierarchy and conformity. We discuss how idealised representations of democracy justify global power arrangements and emphasise the paradoxical justification function of democratic values through which nondemocratic forms of social regulation based on physical force are legitimised with the very democratic norms that call for peaceful resolution of conflicts

Inflammatory endotypes of chronic rhinosinusitis based on cluster analysis of biomarkers

Background: Current phenotyping of chronic rhinosinusitis (CRS) into chronic rhinosinusitis with nasal polyps (CRSwNP) and chronic rhinosinusitis without nasal polyps (CRSsNP) might not adequately reflect the pathophysiologic diversity within patients with CRS. Objective: We sought to identify inflammatory endotypes of CRS. Therefore we aimed to cluster patients with CRS based solely on immune markers in a phenotype-free approach. Secondarily, we aimed to match clusters to phenotypes. Methods: In this multicenter case-control study patients with CRS and control subjects underwent surgery, and tissue was analyzed for IL-5, IFN-gamma, IL-17A, TNF-alpha, IL-22, IL-1 beta, IL-6, IL-8, eosinophilic cationic protein, myeloperoxidase, TGF-beta 1, IgE, Staphylococcus aureus enterotoxin-specific IgE, and albumin. We used partition-based clustering. Results: Clustering of 173 cases resulted in 10 clusters, of which 4 clusters with low or undetectable IL-5, eosinophilic cationic protein, IgE, and albumin concentrations, and 6 clusters with high concentrations of those markers. The group of IL-5-negative clusters, 3 clusters clinically resembled a predominant chronic rhinosinusitis without nasal polyps (CRSsNP) phenotype without increased asthma prevalence, and 1 cluster had a T(H)17 profile and had mixed CRSsNP/CRSwNP. The IL-5-positive clusters were divided into a group with moderate IL-5 concentrations, a mixed CRSsNP/CRSwNP and increased asthma phenotype, and a group with high IL-5 levels, an almost exclusive nasal polyp phenotype with strongly increased asthma prevalence. In the latter group, 2 clusters demonstrated the highest concentrations of IgE and asthma prevalence, with all samples expressing Staphylococcus aureus enterotoxin-specific IgE. Conclusion: Distinct CRS clusters with diverse inflammatory mechanisms largely correlated with phenotypes and further differentiated them and provided a more accurate description of the inflammatory mechanisms involved than phenotype information only

Finding Single Copy Genes Out of Sequenced Genomes for Multilocus Phylogenetics in Non-Model Fungi

Historically, fungal multigene phylogenies have been reconstructed based on a small number of commonly used genes. The availability of complete fungal genomes has given rise to a new wave of model organisms that provide large number of genes potentially useful for building robust gene genealogies. Unfortunately, cross-utilization of these resources to study phylogenetic relationships in the vast majority of non-model fungi (i.e. “orphan” species) remains an unexamined question. To address this problem, we developed a method coupled with a program named “PHYLORPH” (PHYLogenetic markers for ORPHans). The method screens fungal genomic databases (107 fungal genomes fully sequenced) for single copy genes that might be easily transferable and well suited for studies at low taxonomic levels (for example, in species complexes) in non-model fungal species. To maximize the chance to target genes with informative regions, PHYLORPH displays a graphical evaluation system based on the estimation of nucleotide divergence relative to substitution type. The usefulness of this approach was tested by developing markers in four non-model groups of fungal pathogens. For each pathogen considered, 7 to 40% of the 10–15 best candidate genes proposed by PHYLORPH yielded sequencing success. Levels of polymorphism of these genes were compared with those obtained for some genes traditionally used to build fungal phylogenies (e.g. nuclear rDNA, β-tubulin, γ-actin, Elongation factor EF-1α). These genes were ranked among the best-performing ones and resolved accurately taxa relationships in each of the four non-model groups of fungi considered. We envision that PHYLORPH will constitute a useful tool for obtaining new and accurate phylogenetic markers to resolve relationships between closely related non-model fungal species